In silico Cloning of Homogentisate Phytyltransferase Gene from Soybean

doi:10.7668/hbnxb.2007.04.004

Abstract

Abstract: In silico cloning based on EST sequence is a new method of gene cloning, using Arabidopsis thaliana homogentisate phytyltransferase cDNA sequence as a query probe to blast Glycine max EST database, a soybean HPT gene cDNA of 1777 bp was obtained (GeneBank Accession, AY956421). This sequence was confirmed by RT-PCR, molecular cloning and sequencing, It contained a complete ORF, from 173 bp to 1408 bp, encoded 411 amino acids, and conserved with Arabidopsis thaliana, Medicago sative, Oryza, Zea mays and Synechocystis. The results show that it is an efficient technique to clone new genes by searching EST database with homologous gene of model species.

Key words: In silico cloning, EST database, RT-PCR, Homogentisate phytyltransferase, Soybean

CLC Number:

S565.103

MENG Xian-ping, LI Xiao-xiao, LI Ya-xuan, CAI Min-hua, HU Ying-kao. In silico Cloning of Homogentisate Phytyltransferase Gene from Soybean[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2007, 22(4): 14-18. doi: 10.7668/hbnxb.2007.04.004.

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